Work will continue on two projects. First, we propose to characterize those monoclonal antibodies that we have obtained and that are directed against proteins specified by reovirus. This characterization will involve determining their specificity by three methods. First, IgGs against the same protein secreted by several hybridoma clones will be radio-iodinated and the ability of the various unlabled hybridoma IgGs that precipitate the same protein to compete in binding assays with labeled IgG will be determined. Second, we will evaluate the ability of these various IgGs to bind peptides of the proteins with which they react. Third, the various IgGs will be tested for their ability to interact with proteins of the two other serotypes. Other monoclonal antibodies will also be prepared and their specificities determined. We will prepare monoclonal antibodies against the remaining three proteins of reovirus type 3, as well as against protein 1 of types 1 and 2. Monoclonal antibodies will also be prepared against several type-specific and cross-reacting protein of herpes simplex virus types 1 and 2. Second, we will develop techniques employing the monoclonal antibodies to detect very small amounts of specific viral antigens and antibodies. Such techniques will serve two purposes: they can be employed to study viral morphogenesis in infected cells or they can be employed in the early diagnosis of viral disease. Initially the techniques will be modifications of the SPIRA procedure. Eventually the hybridoma IgGs will be linked covalently to alkaline phosphatase (ELISA), as this results in more stable reagents.